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目的研究芳香族氨基酸脱羧酶(AADC)基因和NURR1基因联合c17．2神经干细胞脑内移植后对帕金森病模型大鼠的治疗效果。方法人类神经元性AADC基因和NURR1基因真核表达载体分别转染至c17．2神经干细胞内。将帕金森病(PD)模型大鼠随机分为4组,分别予以脑内毁损侧纹状区移植含空质粒的c17．2神经干细胞(A组),pCDNA3-AADC转染后的c17．2神经干细胞(B组),pCDNA3-NURR1转染后的c17．2神经干细胞(C组)以及含有pCDNA3-AADC和pCDNA3-NURR1转染后的c17．2 神经干细胞(D组)。观察其病理性旋转行为的改善,采用酪氨酸羟化酶(TH)的免疫组化方法研究脑内多巴胺含量的变化,并用荧光示踪方法观察c17．2细胞在PD模型脑内的移行。结果各组动物脑内移植后动物旋转行为较前均有改善(P<0．05),尤以D组改善最为明显,其行为学最大改善达73．7％,且同A、B、C组间差异具有显著性(P<0．05)。免疫组化可见各组移植TH阳性细胞明显增多,TH染色的神经元树突或轴突密集,体内TH阳性区域明显较PD模型组扩大,其中尤以D组病理学改善最为明显。荧光示踪观察c17．2神经干细胞有突触形成,并与临近的细胞建立突触联系。结论 AADC基因联合NURR1基因共转染c17．2神经干细胞脑内移植后改善了动物的旋转行为,增加了脑内多巴胺的表达,且植入的神经干细胞可同宿主神经元形成突触联接,为研究多基因联合神经干细胞移植治疗帕金森病提供了新方法。
Objective To investigate the therapeutic effect of Amino Acids Decarboxylase (AADC) gene and NURR1 gene combined with c17.2 neural stem cells transplantation in Parkinson’s disease model rats. Methods Human neuronal AADC gene and NURR1 gene eukaryotic expression vector were transfected into c17.2 neural stem cells respectively. The Parkinson’s disease (PD) model rats were randomly divided into 4 groups. The c17.2 neural stem cells (A group) containing the empty plasmid were transplanted into the lesioned lateral striatum in the brain. The c17.2 cells transfected with pCDNA3-AADC Neural stem cells (group B), c17.2 neural stem cells transfected with pCDNA3-NURR1 (group C), and c17.2 neural stem cells transfected with pCDNA3-AADC and pCDNA3-NURR1 (group D). To observe the improvement of pathological rotation, the tyrosine hydroxylase (TH) immunohistochemical method was used to study the change of dopamine content in the brain, and the c17.2 cell migration in the PD model was observed by fluorescence tracing method. Results The rotation behavior of the animals after intracerebral transplantation in all groups was improved compared with those before (P <0.05). Especially in group D, the improvement was the most obvious. The behavioral improvement was 73.7% and the same as that of A, B, C Differences between groups were significant (P <0.05). Immunohistochemistry showed that the number of TH-positive cells transplanted in each group was significantly increased. TH-stained neurons dendrites or axons were densely populated. The TH-positive regions in vivo were significantly enlarged compared with PD model group, especially in group D. Fluorescent tracing showed that synapses were formed in c17.2 neural stem cells and synapse contacts were established with adjacent cells. Conclusion AADC gene co-transfected with NURR1 gene translocates c17.2 neural stem cells to brain and improve the animal’s rotational behavior and increase dopamine expression in the brain. The implanted neural stem cells can form synaptic connections with the host neurons The study of multi-gene combined with neural stem cell transplantation for the treatment of Parkinson’s disease provides a new method.